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A Flow Chamber Assay for Studying MAIT Cell Trafficking.
Farhat Parween1, Hongwei H Zhang1, Joshua M Farber2
1Inflammation Biology Section, Laboratory of Molecular Immunology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.
Methods in Molecular Biology (Clifton, N.J.)
|December 4, 2019
Summary
Human MAIT cells traffic to inflamed tissues, not lymph nodes, due to specific receptor expression. This study models MAIT cell extravasation to understand their migration into tissues.
Area of Science:
- Immunology
- Cell Biology
Background:
- Mucosal-AssociatedInvariant T (MAIT) cells are crucial immune cells.
- MAIT cells exhibit distinct surface marker expression, favoring tissue infiltration over lymph node entry.
Purpose of the Study:
- To investigate the molecular mechanisms governing MAIT cell trafficking into inflamed tissues.
- To model and analyze the extravasation process of MAIT cells.
Main Methods:
- Utilized fluorescence-activated cell sorting to purify human MAIT cells from lymphocytes.
- Employed flow chambers with endothelial cell monolayers to simulate leukocyte extravasation under physiological shear stress.
- Assessed the roles of specific chemokine receptors in individual extravasation steps.
Main Results:
- MAIT cells express low levels of lymph node-homing molecules (CD62L, CCR7).
- MAIT cells express high levels of selectin ligands and chemokine receptors facilitating inflamed tissue trafficking.
- Demonstrated the involvement of individual chemokine receptors in specific MAIT cell extravasation steps.
Conclusions:
- MAIT cell trafficking is directed towards inflamed tissues rather than lymph nodes.
- The developed flow chamber model effectively dissects molecular mechanisms of MAIT cell extravasation.
- This research provides a framework for studying MAIT cell migration into tissues.
